Site directed mutagenesis of TREM-1 antibodies for decreasing viscosity

Abstract

Antibodies that are capable of specifically binding and preventing the activation of TREM-1, a protein expressed on monocytes, macrophages and neutrophils with both good affinity and low viscosity at clinically relevant concentrations are described. Such antibodies find utility in the treatment of individuals with an inflammatory disease, such as rheumatoid arthritis and inflammatory bowel disease.

The invention claimed is: 
     
       1. An antibody or fragment thereof (“antibody”) that is capable of binding to and blocking TREM-1, which comprises a light chain CDR1 region, a light chain CDR2 region, and a light chain CDR3 region, a heavy chain CDR1 region, a heavy chain CDR2 region, and a heavy chain CDR3 region,
 wherein the light chain CDR1 region, the light chain CDR2 region, and the light chain CDR3 are the light chain CDR1 region, the light chain CDR2 region, and the light chain CDR3 region present in a light chain selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and SEQ ID NO: 14; and 
 wherein the heavy chain CDR1 region, the heavy chain CDR2 region, and the heavy chain CDR3 are the heavy chain CDR1 region, the heavy chain CDR2 region, and the heavy chain CDR3 region present in a heavy chain selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 15, and SEQ ID NO: 16. 
 
     
     
       2. The antibody of  claim 1 , which comprises a constant region. 
     
     
       3. The antibody of  claim 2 , wherein the constant region is IgG1, IgG2, IgG3, or IgG4. 
     
     
       4. The antibody of  claim 1 , wherein the heavy chain CDR regions are the CDR regions of the heavy chain as set forth in SEQ ID NO: 2. 
     
     
       5. The antibody of  claim 4 , wherein the light chain CDR regions are the CDR regions of the light chain as set forth in SEQ ID NO: 5. 
     
     
       6. The antibody of  claim 5 , which comprises an IgG4 constant region. 
     
     
       7. The antibody of  claim 5 , which comprises a light chain and a heavy chain, wherein the light chain comprises the amino acid sequence as set forth in SEQ ID NO: 5 and the heavy chain comprises the amino acid sequence as set forth in SEQ ID NO: 2. 
     
     
       8. An isolated polynucleotide that encodes the antibody of  claim 1 . 
     
     
       9. A vector comprising the isolated polynucleotide of  claim 8 . 
     
     
       10. A cell comprising the isolated polynucleotide of  claim 8 . 
     
     
       11. A method of making an antibody or fragment thereof which is capable of binding TREM-1, comprising culturing the cell of  claim 10 . 
     
     
       12. An antibody or fragment thereof (“antibody”) that is capable of binding to and blocking TREM-1, which comprises a light chain CDR1 region, a light chain CDR2 region, and a light chain CDR3 region, a heavy chain CDR1 region, a heavy chain CDR2 region, and a heavy chain CDR3 region,
 wherein the light chain CDR1 region, the light chain CDR2 region, and the light chain CDR3 are the light chain CDR1 region, the light chain CDR2 region, and the light chain CDR3 region present in the light chain as set forth in SEQ ID NO: 3; and 
 wherein the heavy chain CDR1 region, the heavy chain CDR2 region, and the heavy chain CDR3 are the heavy chain CDR1 region, the heavy chain CDR2 region, and the heavy chain CDR3 region present in a heavy chain selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 15, and SEQ ID NO: 16. 
 
     
     
       13. The antibody of  claim 12 , which comprises a constant region. 
     
     
       14. The antibody of  claim 13 , wherein the constant region is IgG1, IgG2, or IgG4. 
     
     
       15. An isolated polynucleotide that encodes the antibody of  claim 12 . 
     
     
       16. A cell comprising the isolated polynucleotide of  claim 15 . 
     
     
       17. A method of making an antibody or fragment thereof which is capable of binding TREM-1, comprising culturing the cell of  claim 16 .